title: Development of a high-throughput antiviral assay for screening inhibitors of chikungunya virus and generation of drug-resistant mutations in cultured cells.
abstract: Chikungunya virus (CHIKV) is a mosquito-borne Alphavirus that has already infected millions of people in recent large-scale epidemics in Africa, the islands of the Indian Ocean, South and Southeast Asia, and northern Italy. The infection is still ongoing in many countries, such as India. Although the fatal rate is approximately 0.1% in the La Réunion outbreak, it causes painful arthritis-like symptoms that can last for months or even years. Currently, neither vaccine nor approved antiviral therapy exists to protect humans from chikungunya infection. Therefore, there is an urgent unmet medical need for the development of antiviral drugs for pre-exposure prophylaxis and/or treatment of chikungunya infections. In this chapter, we describe a fully validated ATP/luminescence assay that is effective for high-throughput screening of CHIKV inhibitors. Protocols for growing CHIKV stocks and generating drug-resistant viral variants for modes of action studies of compounds are also described.
authors.affiliation: Janssen Infectious Diseases-Diagnostics BVBA, Johnson & Johnson Corporation, Beerse, Belgium.
authors.forename: Edwin Yunhao
authors.initials: EY
authors.lastname: Gong
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Bonfanti
authors.forename: Tania
authors.initials: T
authors.lastname: Ivens
authors.forename: Marijke
authors.initials: M
authors.lastname: Van der Auwera
authors.forename: Barbara
authors.initials: B
authors.lastname: Van Kerckhove
authors.forename: Guenter
authors.initials: G
authors.lastname: Kraus
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002646:Chikungunya virus / Q000187:drug effects* / Q000235:genetics*; D002522:Chlorocebus aethiops; D024882:Drug Resistance, Viral / Q000235:genetics*; D056945:Hep G2 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009154:Mutation; D014709:Vero Cells
chemical_list: D000998:Antiviral Agents